Antimicrobial Polypeptides Capable of Membrane Translocation for Treatment of MRSA Wound Infection in Vivo

Mitochondrial proteins are synthesized on cytosolic ribosomes and imported into mitochondria with the help of protein translocases. For the majority of precursor proteins, the role of the translocase of the outer membrane (TOM) and mechanisms of their transport across the outer mitochondrial membrane are well recognized. However, little is known about the mode of membrane translocation for proteins that are targeted to the intermembrane space via the redox-driven mitochondrial intermembrane space import and assembly (MIA) pathway. On the basis of the results obtained from an in organello competition import assay, we hypothesized that MIA-dependent precursor proteins use an alternative pathway to cross the outer mitochondrial membrane. Here we demonstrate that this alternative pathway involves the protein channel formed by Tom40. We sought a translocation intermediate by expressing tagged versions of MIA-dependent proteins in vivo. We identified a transient interaction between our model substrates and Tom40. Of interest, outer membrane translocation did not directly involve other core components of the TOM complex, including Tom22. Thus MIA-dependent proteins take another route across the outer mitochondrial membrane that involves Tom40 in a form that is different from the canonical TOM complex.

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Photo-Crosslinked Polymeric Matrix with Antimicrobial Functions for Excisional Wound Healing in Mice

Nanomaterials ◽

10.3390/nano8100791 ◽

2018 ◽

Vol 8 (10) ◽

pp. 791 ◽

Cited By ~ 3

Author(s):

Ming-Hsiang Chang ◽

Yu-Ping Hsiao ◽

Chia-Yen Hsu ◽

Ping-Shan Lai

Keyword(s):

Wound Healing ◽

Wound Infection ◽

Polymer Solution ◽

Wound Dressing ◽

Glycol Diacrylate ◽

Excisional Wound ◽

Poly Ethylene ◽

Systemic Infections

Wound infection extends the duration of wound healing and also causes systemic infections such as sepsis, and, in severe cases, may lead to death. Early prevention of wound infection and its appropriate treatment are important. A photoreactive modified gelatin (GE-BTHE) was synthesized by gelatin and a conjugate formed from the 3,3′,4,4′-benzophenone tetracarboxylic dianhydride (BTDA) and the 2-hydroxyethyl methacrylate (HEMA). Herein, we investigated the photocurable polymer solution (GE-BTHE mixture) containing GE-BTHE, poly(ethylene glycol) diacrylate (PEGDA), chitosan, and methylene blue (MB), with antimicrobial functions and photodynamic antimicrobial chemotherapy for wound dressing. This photocurable polymer solution was found to have fast film-forming property attributed to the photochemical reaction between GE-BTHE and PEGDA, as well as the antibacterial activity in vitro attributed to the ingredients of chitosan and MB. Our in vivo results also demonstrated that untreated wounds after 3 days had the same scab level as the GE-BTHE mixture-treated wounds after 20 s of irradiation, which indicates that the irradiated GE-BTHE mixture can be quickly transferred into artificial scabs to protect wounds from an infection that can serve as a convenient excisional wound dressing with antibacterial efficacy. Therefore, it has the potential to treat nonhealing wounds, deep burns, diabetic ulcers and a variety of mucosal wounds.

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Sanguiin H-6 Fractionated from Cloudberry (Rubus chamaemorus) Seeds Can Prevent the Methicillin-Resistant Staphylococcus aureus Biofilm Development during Wound Infection

Antibiotics ◽

10.3390/antibiotics10121481 ◽

2021 ◽

Vol 10 (12) ◽

pp. 1481

Author(s):

John Jairo Aguilera-Correa ◽

Sara Fernández-López ◽

Iskra Dennisse Cuñas-Figueroa ◽

Sandra Pérez-Rial ◽

Hanna-Leena Alakomi ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Wound Infection ◽

Preventive Measure ◽

Surgical Site Infections ◽

Infection Model ◽

Growth Media ◽

Methicillin Resistant ◽

Biofilm Development

Staphylococcus aureus is the most common cause of surgical site infections and its treatment is challenging due to the emergence of multi-drug resistant strains such as methicillin-resistant S. aureus (MRSA). Natural berry-derived compounds have shown antimicrobial potential, e.g., ellagitannins such as sanguiin H-6 and lambertianin C, the main phenolic compounds in Rubus seeds, have shown antimicrobial activity. The aim of this study was to evaluate the effect of sanguiin H-6 and lambertianin C fractionated from cloudberry seeds, on the MRSA growth, and as treatment of a MRSA biofilm development in different growth media in vitro and in vivo by using a murine wound infection model where sanguiin H-6 and lambertianin C were used to prevent the MRSA infection. Sanguiin H-6 and lambertianin C inhibited the in vitro biofilm development and growth of MRSA. Furthermore, sanguiin H-6 showed significant anti-MRSA effect in the in vivo wound model. Our study shows the possible use of sanguiin H-6 as a preventive measure in surgical sites to avoid postoperative infections, whilst lambertianin C showed no anti-MRSA activity.

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Nontoxic membrane translocation peptide from protamine, low molecular weight protamine (LMWP), for enhanced intracellular protein delivery: in vitro and in vivo study

The FASEB Journal ◽

10.1096/fj.04-2322fje ◽

2005 ◽

Vol 19 (11) ◽

pp. 1555-1557 ◽

Cited By ~ 85

Author(s):

Yoon Jeong Park ◽

Li‐Chien Chang ◽

Jun Feng Liang ◽

Cheol Moon ◽

Chong‐Pyoung Chung ◽

...

Keyword(s):

Molecular Weight ◽

Protein Delivery ◽

In Vivo Study ◽

Low Molecular Weight ◽

Membrane Translocation ◽

Intracellular Protein ◽

Intracellular Protein Delivery

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Pentraxin 3 protects from MCMV infection and reactivation through TLR sensing pathways leading to IRF3 activation

Blood ◽

10.1182/blood-2006-03-009266 ◽

2006 ◽

Vol 108 (10) ◽

pp. 3387-3396 ◽

Cited By ~ 96

Author(s):

Silvia Bozza ◽

Francesco Bistoni ◽

Roberta Gaziano ◽

Lucia Pitzurra ◽

Teresa Zelante ◽

...

Keyword(s):

Viral Entry ◽

Allogeneic Transplantation ◽

Interleukin 12 ◽

Viral Fusion ◽

Pentraxin 3 ◽

Mcmv Infection ◽

Antimicrobial Polypeptides ◽

The Impact

AbstractReactivation of latent human cytomegalovirus (HCMV) following allogeneic transplantation is a major cause of morbidity and mortality and predisposes to severe complications, including superinfection by Aspergillus species (spp). Antimicrobial polypeptides, including defensins and mannan-binding lectin, are known to block viral fusion by cross-linking sugars on cell surface. Pentraxin 3 (PTX3), a member of the long pentraxin family, successfully restored antifungal immunity in experimental hematopoietic transplantation. We assessed here whether PTX3 binds HCMV and murine virus (MCMV) and the impact on viral infectivity and superinfection in vivo. We found that PTX3 bound both viruses, reduced viral entry and infectivity in vitro, and protected from MCMV primary infection and reactivation as well as Aspergillus superinfection. This occurred through the activation of interferon (IFN) regulatory factor 3 (IRF3) in dendritic cells via the TLR9/MyD88-independent viral recognition sensing and the promotion of the interleukin-12 (IL-12)/IFN-γ–dependent effector pathway.

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Organo-Selenium-Containing Polyester Bandage Inhibits Bacterial Biofilm Growth on the Bandage and in the Wound

Biomedicines ◽

10.3390/biomedicines8030062 ◽

2020 ◽

Vol 8 (3) ◽

pp. 62

Author(s):

Phat Tran ◽

Tyler Enos ◽

Keaton Luth ◽

Abdul Hamood ◽

Coby Ray ◽

...

Keyword(s):

Wound Infection ◽

Wound Dressing ◽

Laser Scanning ◽

Bacterial Biofilm ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Bacterial Strains ◽

Biofilm Growth ◽

Biofilm Model

The dressing material of a wound plays a key role since bacteria can live in the bandage and keep re-infecting the wound, thus a bandage is needed that blocks biofilm in the bandage. Using an in vivo wound biofilm model, we examined the effectiveness of an organo-selenium (OS)-coated polyester dressing to inhibit the growth of bacteria in a wound. Staphylococcus aureus (as well as MRSA, Methicillin resistant Staph aureus), Stenotrophomonas maltophilia, Enterococcus faecalis, Staphylococcus epidermidis, and Pseudomonas aeruginosa were chosen for the wound infection study. All the bacteria were enumerated in the wound dressing and in the wound tissue under the dressing. Using colony-forming unit (CFU) assays, over 7 logs of inhibition (100%) was found for all the bacterial strains on the material of the OS-coated wound dressing and in the tissue under that dressing. Confocal laser scanning microscopy along with IVIS spectrum in vivo imaging confirmed the CFU results. Thus, the dressing acts as a reservoir for a biofilm, which causes wound infection. The same results were obtained after soaking the dressing in PBS at 37 °C for three months before use. These results suggest that an OS coating on polyester dressing is both effective and durable in blocking wound infection.

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2477

Journal of Clinical and Translational Science ◽

10.1017/cts.2017.233 ◽

2017 ◽

Vol 1 (S1) ◽

pp. 65-65

Author(s):

Petra Wilder-Smith ◽

Janet Ajdaharian ◽

Afarin Golabgir Anbarani ◽

Jessica Ho ◽

Karan Sahni ◽

...

Keyword(s):

Wound Infection ◽

Imaging Modality ◽

Focal Length ◽

Wound Response ◽

Bacterial Biofilm ◽

The Novel ◽

Susceptibility To Infection ◽

Micro Channels ◽

Biofilm Infection

OBJECTIVES/SPECIFIC AIMS: The specific objectives of this project are (1) identify, test, and validate the parameters for a simplified NLOM imaging probe that will provide specific research and point-of-care information on biofilm presence, therapeutic need and response of individual wounds to treatment. (2) Identify specific proteomic and metabolomic biomarkers of (i) wound susceptibility to infection, (ii) wound response to the most commonly used antibacterial measures in wounds, and (iii) establish criteria for more effective interventions. METHODS/STUDY POPULATION: First, optimal use parameters for NLOM including illumination, field of view, focal length, linear Versus concentric image acquisition, detection and filter wavelengths were identified. Parameters for evaluation included ease and speed of imaging, ability to map diagnostic criteria. Next, using the optimised NLOM imaging modality in bacterial biofilm isolates and subsequently a rabbit ear model of biofilm wound infection, proteomic and metabolomic biomarkers of susceptibility to infection were identified. The effects of 2 standard debridement and anti-infective treatments, polyvidone-iodine solution or cetrimide 15%+ chlorhexidine gluconate 1.5% were mapped in situ for up to 10 days using the NLOM probe. RESULTS/ANTICIPATED RESULTS: Using the novel custom NLOM probe, high resolution mapping of wound biofilm infection, as well as the underlying tissue was performed throughout the onset, development, treatment, and resolution of wound biofilm infection. Specific microbiological, microstructural, oxygenation, and pH parameters were mapped at defined surface and subsurface locations and time-points. Findings included the determination that some standard antimicrobial formulations provide a supportive environment for wound infection, and that micro-channels within the biofilm and their interface with the tissues serve as an important predictor and indicator of wound infection establishment, progression, and response. DISCUSSION/SIGNIFICANCE OF IMPACT: The novel multimodality in vivo NLOM imaging approach establishes an important tool for earlier and more specific diagnosis of wound infection risk, virulence, and invasiveness along with markers of successful treatment, and a simple clinical imaging tool for improving wound infection prevention and treatment.

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Enterococcus faecalis persists and replicates within epithelial cells in vitro and in vivo during wound infection

10.1101/2021.09.16.460717 ◽

2021 ◽

Author(s):

Wei Hong Tay ◽

Ronni A.G. da Silva ◽

Foo Kiong Ho ◽

Kelvin K.L. Chong ◽

Alexander Ludwig ◽

...

Keyword(s):

Wound Infection ◽

Enterococcus Faecalis ◽

Opportunistic Pathogen ◽

Small Gtpase ◽

Infection Model ◽

Robust Immune Response ◽

Lysosomal Protease ◽

The Face

Enterococcus faecalis is a frequent opportunistic pathogen of wounds, whose infections are associated with biofilm formation, persistence, and recalcitrance toward treatment. We have previously shown that E. faecalis wound infection persists for at least 7 days. Here we report that viable E. faecalis are present within both immune and non-immune cells at the wound site up to 5 days after infection, raising the prospect that intracellular persistence contributes to chronic E. faecalis infection. Using an in vitro keratinocyte infection model, we show that a subpopulation of E. faecalis becomes internalized via macropinocytosis into single membrane-bound compartments, where they can survive and replicate. These intracellular E. faecalis can persist in late endosomes up to 72 hours after infection in the absence of colocalization with the lysosomal protease cathepsin D or apparent fusion with the lysosome, suggesting that E. faecalis blocks endosomal maturation. Indeed, intracellular E. faecalis infection results in a marked reduction in Rab7 expression, a small GTPase required for endosome-lysosome fusion. Finally, we demonstrate that intracellular E. faecalis derived from infected keratinocytes are significantly more efficient in reinfecting new keratinocytes. Together, these data suggest that intracellular proliferation of E. faecalis may contribute to its persistence in the face of a robust immune response, providing a primed reservoir of bacteria for subsequent reinfection.

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Challenges with Wound Infection Models in Drug Development

Current Drug Targets ◽

10.2174/1389450121666200302093312 ◽

2020 ◽

Vol 21 (13) ◽

pp. 1301-1312 ◽

Cited By ~ 1

Author(s):

Sandeep K. Shukla ◽

Ajay K. Sharma ◽

Vanya Gupta ◽

Aman Kalonia ◽

Priyanka Shaw

Keyword(s):

Wound Healing ◽

Wound Infection ◽

Chronic Wound ◽

Wound Care ◽

Infection Model ◽

In Vivo Models ◽

Infection Models

: Wound research is an evolving science trying to unfold the complex untold mechanisms behind the wound healing cascade. In particular, interest is growing regarding the role of microorganisms in both acute and chronic wound healing. Microbial burden plays an important role in the persistence of chronic wounds, ultimately resulting in delayed wound healing. It is therefore important for clinicians to understand the evolution of infection science and its various etiologies. Therefore, to understand the role of bacterial biofilm in chronic wound pathogenesis, various in vitro and in vivo models are required to investigate biofilms in wound-like settings. Infection models should be refined comprising an important signet of biofilms. These models are eminent for translational research to obtain data for designing an improved wound care formulation. However, all the existing models possess limitations and do not fit properly in the model frame for developing wound care agents. Among various impediments, one of the major drawbacks of such models is that the wound they possess does not mimic the wound a human develops. Therefore, a novel wound infection model is required which can imitate the human wounds. : This review article mainly discusses various in vitro and in vivo models showing microbial colonization, their advantages and challenges. Apart from these models, there are also present ex vivo wound infection models, but this review mainly focused on various in vitro and in vivo models available for studying wound infection in controlled conditions. This information might be useful in designing an ideal wound infection model for developing an effective wound healing formulation.

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